Fig. 5: The relationship between autophagy and JA pathway and the role of ERF1.
From: Autophagy promotes jasmonate-mediated defense against nematodes
a, b Content of JA (a) and JA-Ile (b) in Ailsa Craig (AC), atg7, Castlemart (CM) and spr2 mutants at 48 hpi with RKN. JA jasmonic acid, JA-Ile jasmonoyl-isoleucine. Error bars represent SD; data represent the mean ± SD (n = 3 biological replicates, individual dots). c Gene expression of ERF1 in AC, atg7, CM and spr2 mutants at 48 hpi with RKNs. Error bars represent SD; data represent the mean ± SD (n = 3 biological replicates, individual dots). d Protein levels of ERF1 in AC, atg7, CM and spr2 mutants at 48 hpi with RKNs. The anti-ERF1 antibody was used to detect the protein level of ERF1 and the Actin protein was used as a loading control. The experiments were repeated twice with similar results. e Electrophoretic mobility shift assays to test His-ERF1 recombinant proteins bound to the probe from the ERF1 promoter harboring the key GCCGCC (GCC-box) motif (probe-ERF1). Mu, mutated probe in which the GCCGCC motif was changed to AAAAAA. Competitor and mutant competitor (Mu-competitor) were used at 1000-fold. The experiments were repeated twice with similar results. f Chromatin immunoprecipitation (ChIP)-real-time quantitative PCR assays to test direct binding of ERF1 to its self-promoter with RKN infection. The epitope-tagged protein-chromatin complexes were immunoprecipitated with an anti-GFP antibody (Ab). Control reactions were treated with mouse immunoglobulin G (IgG). Error bars represent SD; data represent the mean ± SD (n = 3 biological replicates, individual dots). g Regulatory effects of ERF1 on the promoter of ERF1 as determined by dual-luciferase assays. The ratios of firefly luciferase/Renilla luciferase (LUC/REN) of the empty vector (EV) plus promoters under normal conditions were set as ‘1’. Error bars represent SD; data represent the mean ± SD (n = 4 biological replicates). Different letters above bars indicate a significant difference at the P < 0.05 level one-way ANOVA with Tukey’s multiple comparisons test (a–c, f). The asterisks in (g) indicate significant difference as assessed by two-sided Student’s t-tests; *** P < 0.001. Exact P-values of statistic tests are provided in the Source data file.
